<p><strong>Introducing</strong>: ONI’s Next-Generation End-to-End Solution for EV Research <span style="text-decoration: underline;">Learn More</span></p>

Introducing: ONI’s Next-Generation End-to-End Solution for EV Research Learn More

Application Kit™: EV Profiler

Characterize populations of single EVs with the Nanoimager

Harness the power of super-resolution microscopy to characterize EV populations and individual EVs. Quantify biomarkers on small, rare, and precious EVs with single molecule sensitivity. ONI’s EV Profiler helps you focus on answering questions about your EVs.

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Identify EVs and quantify surface biomarkers

With the EV Profiler Kit you use our validated antibodies to measure the expression of tetraspanins and characterize the single, double and triple positive populations, or you can measure your own biomarker of interest.*

Single molecule localization microscopy used in combination with the appropriate fluorescently labeled probes, helps confirm EV identity after purification. ONI’s EV Profiler Kit in combination with cluster-based analysis on our CODI cloud platform, allows reporting of morphological features of the EV, including size and circularity.

*Reagents have a shelf life of two months.

ONI’s EV Profiler Kit allows detection of up to 3 biomarkers on captured EVs. HCT116-derived EV sample were captured with ONI’s S4 molecule, labeled with anti-CD9-CF®488 (yellow), anti-CD63-CF®568 (cyan) and anti-CD81-CF®647 (magenta) and imaged using dSTORM on the Nanoimager (left). Biomarker positivity was quantified using ONI’s CODI cluster-based algorithm on single-molecule localization microscopy data (right).

Flexible Capture of your EVs

EV Profiler enables the capture of EVs on a passivated glass surface. The kit’s workflow is compatible with the following capture methods:

Validated by ONI – to be selected during purchase:

  • Phosphatidylserine-dependent, protein-independent capture
    Our proprietary S4 capture molecule binds to phosphatidylserine providing a capture method that is agnostic to the presence of protein biomarkers on the EVs.
  • Tetraspanin-based capture
    ONI validated capture antibodies against tetraspanins: anti-CD9, anti-CD63 and anti-CD81 antibodies can be purchased separately.

Choose your own:

  • Custom target based capture
    Use your own antibody of choice against your target of interest – it simply needs to be biotinylated. Get in touch with our team to find out more on how to do this.

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Tetraspanin based capture

ONI validated capture antibodies against tetraspanins: anti-CD9, anti-CD63 and anti-CD81 antibodies can be selected when purchasing the kit.

Phosphatidylserine-dependent, protein-independent capture

Our proprietary S4 capture molecule binds EVs in a protein-independent manner, working best with phosphatidylserine-rich EVs.

Custom target based capture

Use your own antibody of choice against your target of interest – it simply needs to be biotinylated (sample above provided by Dr Nicole Meisner-Kober’s lab).

EV characterization across scales: from EV populations down to single EVs

Study EV populations across scales. Each EV Profiler kit contains enough material to analyze 12 EV samples using our 4 lane analysis chip which has been developed and optimized for EV capture. You can capture and image hundreds of EVs from any given source with the appropriate capture method (read more details below), then zoom into individual EVs to see biomarker distribution at a single-EV level.

ONI’s Nanoimager and CODI platform enable imaging and visual inspection of EV samples across scales. Scale bars range from 10 µm to 50 nm when inspecting one EV out of thousands of EVs imaged on from an HCT116-derived EV sample captured with ONI’s S4 molecule and labeled with pan-EV stain (yellow), anti-CD63-CF®568 (cyan) and anti-CD81-CF®647 (magenta).

Obtain rapid results with a benchtop assay

Capture, stain, image and quantify

The EV Profiler Kit contains all necessary reagents to capture, fix and stain EVs ready for imaging and the workflow works with EV sample volumes as low as 10 µL.

Single molecule localization data and nanoscale resolution images can be obtained using the Nanoimager, our benchtop super-resolution microscope. Data is then analyzed through our CODI cloud-based software, making it possible to get results within 24 hours from start to finish.

Flexibility to suit your EV samples

Besides the flexibility that EV Profiler offers for different EV capture methods, you can easily swap the fluorescent antibodies for surface marker detection for your own.

In the EV Profiler Kit, antibodies for standard tetraspanin markers (CD9, CD63, and CD81) are included conjugated to three different fluorophores so you can profile these markers. However, the kit is modular and antibodies can be swapped for your own validated antibodies against targets of interest inside or outside EVs. These need to be conjugated to dSTORM-compatible dyes.

You can find more information on fluorophores for dSTORM imaging here. The kit’s sample preparation protocol will guide you through the steps for incorporating your antibodies into the experiment and our image acquisition and analysis guide will provide you with the information needed when optimizing your antibodies.

EV analysis and quantification made simple

Purchase of the kit includes access to purpose built analysis tools within our CODI cloud platform, which allow seamless processing of single molecule localization data to obtain output summary data files.

  • Seamless upload data of acquired data to CODI cloud platform 
  • Visualization of individual EVs and their biomarkers with the nanoscale resolution
  • Analysis tools tailored to EV workflow:
    • Correction of spatial drift
    • Filtering of individual localizations
    • Cluster-based single molecule 
  • Extract quantitative data on EV populations based on raw data outputs (.csv format):
    • Biomarker positivity per sample
    • EV size distribution
    • Biomarker density per EV (localizations/cluster)
    • EV morphology parameters
  • Export high-resolution 3-color images from your EV samples 

ONI’s CODI cloud software uses clustering of single molecule data to enable study of EVs. Left image shows EVs captured using the EV Profiler Kit, which were imaged with the Nanoimager. Single molecule localization data is then clustered on CODI to help assess EV morphology features and determine levels of positivity for different biomarkers.

EV Profiler is powered by ONI’s Nanoimager and CODI platform

Combine the advantages of super-resolution imaging with powerful analysis of larger numbers of events. This allows detection and quantification of biomarker abundance with up to 3 biomarkers detected within the same sample using dSTORM, and reporting of EV morphology, including size and circularity. 

Whats in the box?

3 four lane flow-cell chips
6 lane stickers
2 stage securing magnets
Capture reagents (3 to choose from)
Phosphatidylserine-dependent S4 capture
Biotinylated anti-CD9 capture
Biotinylated anti-CD63 capture
Biotinylated anti-CD81 capture
Detection probes
Anti-CD9 488 (1 tube)
Anti-CD63 568 (1 tube)
Anti-CD81 647 (1 tube)
E1 standard EVs (1 tube)
S3, W1, F1, B1 (3 tubes each)
C1 (1 tube)
BCubed imaging buffer parts A and B (3 tubes each)

Request more information

Contact one of our sales specialists to request more information or to order EV Profiler V2